CN117701666A - Application of iron death inducer - Google Patents
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- CN117701666A CN117701666A CN202311462585.4A CN202311462585A CN117701666A CN 117701666 A CN117701666 A CN 117701666A CN 202311462585 A CN202311462585 A CN 202311462585A CN 117701666 A CN117701666 A CN 117701666A
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- iron death
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- colon tumor
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- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 title claims abstract description 131
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- 230000034994 death Effects 0.000 title claims abstract description 56
- 239000000411 inducer Substances 0.000 title claims abstract description 41
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- NDLPOXTZKUMGOV-UHFFFAOYSA-N oxo(oxoferriooxy)iron hydrate Chemical compound O.O=[Fe]O[Fe]=O NDLPOXTZKUMGOV-UHFFFAOYSA-N 0.000 description 2
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- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
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- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
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- FNEZBBILNYNQGC-UHFFFAOYSA-N methyl 2-(3,6-diamino-9h-xanthen-9-yl)benzoate Chemical compound COC(=O)C1=CC=CC=C1C1C2=CC=C(N)C=C2OC2=CC(N)=CC=C21 FNEZBBILNYNQGC-UHFFFAOYSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
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- C12N2503/00—Use of cells in diagnostics
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Abstract
The invention provides an application of an iron death inducer, which comprises the following steps: adding an iron death inducer into a target sample, and simultaneously carrying out ultrasonic treatment on the target sample by adopting ultrasonic waves, wherein the target sample is in-vitro tissue or cell suspension, and the tissue or cell comprises colon tumor cells CT-26 or HT-29; the iron death inducer is nano iron oxide particle FMT and is used for providing an iron-rich environment for a target sample; the ultrasound is used for providing a warm environment for a target sample, and compared with the existing iron death inducer, the iron death inducer prepared by the invention can be effectively applied to inhibiting colon tumor.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to application of an iron death inducer.
Background
Iron death is a different way of cell death than apoptosis, necrosis, because iron-dependent lipid peroxide accumulation causes redox imbalance, intracellular lipid peroxide (ROS) reaches up to a level of death, which in turn triggers cell death. In recent years, a great deal of researches show that the iron death plays a role in resisting tumors, an iron death-related regulation mechanism can effectively inhibit the occurrence and development of tumors (research progress Yang Xinhua on the role of iron death in tumor metastasis), and an iron death inducer is expected to become a new strategy for clinically treating tumors.
Nano iron oxide particles FMT (ferumoxytol) are an important iron-based chemical reagent and are mostly used for the treatment of iron deficiency anemia and the radiography of disease diagnosis clinically. The existing anti-tumor effect of FMT alone as a therapeutic agent is not ideal, and although the anti-tumor effect can be improved by combining the FMT with other tumor therapies, the complexity of the therapies greatly discounts the feasibility of clinical application; and FMT is used in relatively high dosage, which may bring about systemic toxic and side effects. In order to reduce the dosage and improve the tumor treatment effect, researchers try to combine FMT with chemotherapy, phototherapy or immunotherapy, and complicated treatment mechanisms and processes limit the application of nano-ferric oxide particles in clinical transformation of tumor treatment.
Ultrasound is a technique commonly used in clinic, and is most widely used in disease treatment, namely high-intensity focused ultrasound and low-intensity pulsed ultrasound. The high-intensity focused ultrasound technology mainly utilizes the heat generation effect of ultrasonic waves to concentrate the energy of the ultrasonic waves to a certain limited area, and further enhances the heat generation effect of the ultrasonic waves to further rapidly raise the temperature of local tissues and destroy the structure or function of pathological tissues, so that the high-intensity focused ultrasound technology is currently used for the thermal ablation treatment of various proliferative or neoplastic diseases of the whole body; the low-intensity pulse ultrasound is mainly used for assisting or rehabilitation treatment of non-neoplastic diseases by utilizing the mechanical effect and cavitation effect of the ultrasound.
Therefore, finding an application method of an iron death inducer with low dosage, simple method and good effect is still a major problem to be solved.
Disclosure of Invention
In view of the above, the invention provides an application of an iron death inducer, which solves the technical problems of high dosage, poor effect and harsh preparation conditions of the iron agent in the preparation of the existing iron death inducer.
The technical scheme of the invention is realized as follows:
in one aspect, the invention provides an application of an iron death inducer, comprising the following steps:
adding an iron death inducer into a target sample, simultaneously carrying out ultrasonic treatment on the target sample by adopting ultrasonic waves, controlling the sound intensity and frequency of the ultrasonic waves, maintaining the temperature of the target sample at 43 ℃, and inducing the iron death of the target sample;
the target sample is an isolated tissue or a cell, the cell comprising a colon tumor cell; the iron death inducer is FMT, and is used for providing an iron environment for the target sample; the ultrasound is used to provide a thermal environment for the target sample.
Further preferably, the concentration of FMT is 0.75-1mg/mL based on this regimen.
Further preferably, the molecular weight of the FMT is 750kDa on the basis of this protocol.
On the basis of the scheme, it is further preferable that the sound intensity of the ultrasonic wave is 0.75-1.2W/cm 2 The ultrasonic frequency was 1.0MHz.
Further preferably, on the basis of the scheme, the ultrasonic treatment time of the target sample is 5-10min.
Further preferably, on the basis of this protocol, the culture conditions of the target sample are 37℃and 5% CO 2 An incubator.
Further preferably, on the basis of this protocol, the colon tumor cells are CT-26 or HT-29 cells.
The application of the iron death inducer has the following beneficial effects compared with the prior art:
the application of the existing iron death inducer to the inhibition of tumor cells does not appear, the combination of FMT and ultrasound has not occurred, the anticancer effect of FMT alone used as a therapeutic agent is not ideal, and the FMT has relatively high dosage and may bring about some systemic toxic and side effects; the tumor tissue cells are treated by ultrasound, the sound wave intensity is properly regulated, so that the tumor tissue is moderately heated, and the Fenton reaction can be effectively accelerated at 43 ℃ by providing iron and heat, so that the tumor cells die and die, and the treatment of tumors is realized.
The invention realizes the tumor inhibition effect of FMT through ultrasonic synergy on one hand, and provides new disease indication for FMT on the other hand.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing fluorescence intensity according to examples 1-3 of the present invention;
FIG. 2 is a fluorescent image of CD4 and CD8 expression in subcutaneous colon tumor tissue of the mice of example 1 and comparative examples 1-3 of the present invention;
FIG. 3 shows the expression of CD4 and CD8 in subcutaneous colon tumor tissue of the mice of example 1 and comparative examples 1-3;
FIG. 4 shows the change in subcutaneous colon tumor volume after treatment of mice of example 1 and comparative examples 1-3 of the present invention;
FIG. 5 is a fluorescent image of GPX4 expression in subcutaneous colon tumor cells of the mice of example 1 and comparative examples 1-3 of the present invention;
FIG. 6 shows the expression of GPX4 in subcutaneous colon tumor cells of the mice of example 1 and comparative examples 1-3 of the present invention.
Detailed Description
The following description of the embodiments of the present invention will clearly and fully describe the technical aspects of the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, are intended to fall within the scope of the present invention.
The invention provides an application of an iron death inducer, and in a preferred embodiment, the iron death inducer adopts the following materials and instruments:
the CAS number of the FMT is 722492-56-0; the tumor cells comprise mouse colon cancer cells CT26 and human colon cancer cells HT-29 which are purchased from a cell culture library in the United states, and the human peripheral blood mononuclear cells are purchased from Miaotong biotechnology Co., ltd; the BALB/c female mice (4-6 weeks) were supplied by GemPharmatech (south tokyo, china), animals were kept under specific pathogen-free conditions and food and water were freely available.
Cell counting kit-8 (CCK-8) and 4', 6-diamino-2-phenylindole provided by Beyotime Biotechnology, BODIPY TM 665/676 is selected from the group consisting of Siemens' Feishan science, anti-mouse antibodies, goat anti-mouse IgG for immunostaining, GPX4 monoclonal antibodies obtained from Proteintech, anti-mouse antibodies (anti-CD 4, anti-CD 8) and anti-human antibodies (anti-CD 4, anti-CD 8) for flow cytometry analysis were all purchased from BioLegend, hydrogen peroxide (H) 2 O 2 3%) solution and dihydro-hodamine 123 (DHR 123) were from aladin corporation, BD matrix was introduced by corning corporation and the luciferase substrate d-luciferin was purchased from scientific.
Example 1
The present embodiment provides an application of an iron death inducer in inhibiting tumor cells, wherein FMT with a molecular weight of 750kDa is used as the iron death inducer, and specifically comprises the following steps:
s1, establishing a colon tumor model, subcutaneously inoculating colon tumor cells on the right side of a 4-week BALB/c female mouse, and culturing for 5 days; after the BALB/c female mice are weighed, the random groups are designed in groups, each group has 4 mice in weight in each interval, the individual biological difference among the groups of each embodiment is ensured to be minimized, and the consistency principle and the random control principle are followed; CT-26 cells were cultured in RPMI 1640 medium containing 10% FBS and 1% penicillin and under cell culture conditions of 37deg.C and 5% CO 2 An incubator.
S2, 200 mu L of iron death inducer is injected into the tail of the mouse intravenously, the injection dosage is 6mg/kg, the ultrasonic sound intensity is regulated, the ultrasonic sound intensity is monitored by using a thermal imager, and the ultrasonic probe is used for detecting the iron death inducerThe heat of the tumor part is generated at 43 ℃, the ultrasonic frequency is 1.0MHz, and the ultrasonic sound intensity is 0.75W/cm 2 Ultrasonic treatment for 10min; the iron death inducer is FMT and is used for providing iron environment for a target sample; the ultrasound is used to provide a thermal environment for the target sample.
S3, in-vivo tumor imaging, namely injecting 150mg of luciferase substrate into the abdominal cavity of the mice after 7 days of treatment, performing in-vivo bioluminescence imaging, and obtaining a luminescence image through a spectrum imaging system Perkinelmer.
Example 2
This example provides an application of an iron death inducer to tumor cell inhibition, which is different from example 1 in that heat is generated from HT-29 tumor cells by an ultrasonic probe, wherein the heat generation temperature is 23℃and the ultrasonic sound intensity is 1.2W/cm 2 The ultrasonic treatment time is 5min.
Example 3
The present example provides an application of iron death inducer in inhibiting tumor cells, which is different from example 1 in that heat is generated to tumor cells by an ultrasonic probe at 37℃and the ultrasonic sound intensity is 1W/cm 2 The ultrasonic treatment time is 8min.
Wherein the immunofluorescence detection specifically comprises the following steps:
CT-26 cells were inoculated into a confocal chamber at a density of 40000 cells/chamber, incubated at 37℃for 24h, each well of medium was replaced with 1mg/mL of FMT solution, and cells treated with equal volumes of PBS were used as controls and incubated for 24h at a sound intensity of 0.75W/cm 2 Sonicating for 10min, raising the temperature of the culture medium to 43deg.C, maintaining for 10min, culturing at 37deg.C for 12h, washing with 1 times PBS for 3 times, fixing with 4% Paraformaldehyde (PFA) for 10min, and sealing with 3% Bovine Serum Albumin (BSA) at room temperature for 1h; next, stained with a specific antibody to mouse GPX4 (1:400, monoclonal, 67763-1) overnight at 4℃and then stained with Alexa Fluor 488-conjugated goat anti-mouse IgG (1:500, ab 150113) at 25℃for 2h, nuclei stained with 4', 6-diamino-2-phenylindole (DAPI, C1006) and stained cells were visualized using a Zeiss LSM980 confocal fluorescence microscope.
Treating tumor-bearing mice, taking spleen and peripheral blood cells of the mice after 21 days, and performing flow cytometry analysis, wherein the method specifically comprises the following steps:
peripheral blood, spleen, lymph nodes and ascites were isolated from mice and resuspended in PBS and analyzed by labelling immune cells with the corresponding antibodies according to the manufacturer's instructions using anti-mouse antibodies such as CD4 (clone GK1.5, biolgend), CD8 (clone 53-6.7, biolgend).
Monitoring colon tumor growth in mice, comprising: the subcutaneous tumor tissue volumes of the mice of each example were measured daily after treatment, and the tumor volume calculation formula was: volume = tumor length x tumor width 2 /2。
Results: FIG. 1 is a graph showing the fluorescence intensity of ROS expression at various temperatures, and from FIG. 1, it can be seen that DHR123 is used as an index of active oxygen, and the fluorescence intensity of example 1 is significantly higher than that of examples 2 and 3 at various times, indicating that the Fenton reaction rate at 43℃is significantly better than that at 23℃and 37 ℃.
After different treatments, the fluorescence intensities of the colon tumor tissue immune cells CD4 and CD8 of the mice in the example 1 are higher, as shown in figure 1, and after 2-3 days of the treatment, the colon tumor volume of the mice in the example 1 is not obviously changed; after 20 days of treatment, the colon tumor volume of the mice in the example 1 is obviously reduced, which indicates that colon tumor cells are effectively inhibited, and shows that the iron death inducer prepared in the example accelerates Fenton reaction in an iron-rich environment provided by nano ferric oxide under the condition of moderate heating (43 ℃) assisted by ultrasound, so that tumor cells sensitize iron death and apoptosis, thereby exciting immune reaction to overcome inherent immunosuppression tumor microenvironment and being well applied to inhibiting colon tumor cells.
Comparative example 1
The difference from example 1 is that FMT was not added, ultrasound was not performed, and PBS buffer solution commonly used in the prior art was added, specifically comprising the steps of:
s1, establishing a colon tumor model, subcutaneously inoculating colon tumor cells on the right side of a 4-week BALB/c female mouse, and culturing for 5 days; after the BALB/c female mice are weighed, the random groups are designed into groups, and each group has each interval4 mice are weighted, so that the individual biological difference among all the embodiment groups is reduced to the minimum, and the consistency principle and the random control principle are followed; CT-26 cells were cultured in RPMI 1640 medium containing 10% FBS and 1% penicillin and under cell culture conditions of 37deg.C and 5% CO 2 An incubator.
S2, 200 mu L of PBS buffer solution is injected into the tail of the mouse by intravenous injection.
S3, in vivo tumor imaging, injecting 150mg of luciferase substrate into the abdominal cavity of the mouse after 7 days of treatment, performing in vivo bioluminescence imaging, obtaining a luminescent image through a spectrum imaging system Perkinelmer, detecting CT-26 cells, taking spleen and peripheral blood cells of the mouse after 21 days, performing flow cytometry analysis, and monitoring colon tumor growth condition of the mouse.
Results: as shown in fig. 2, after different treatments, the colon tumor tissue of the mice of the control example 1 has low fluorescence intensity, namely, immune cells CD4 and CD8 are few, as shown in fig. 3, after 2-3 days of treatment, the colon tumor volume of the mice of the control example 1 has no obvious change, and after 20 days of treatment, the colon tumor volume of the mice of the control example 1 is increased, which indicates that colon tumor cells are not effectively inhibited.
Comparative example 2
The present embodiment provides an application of an iron death inducer in inhibiting tumor cells, which is different from embodiment 1 in that step S2 is not subjected to ultrasonic treatment, and specifically includes the following steps:
s1, establishing a colon tumor model, subcutaneously inoculating colon tumor cells on the right side of a 4-week BALB/c female mouse, and culturing for 5 days; after the BALB/c female mice are weighed, the random groups are designed in groups, each group has 4 mice in weight in each interval, the individual biological difference among the groups of each embodiment is ensured to be minimized, and the consistency principle and the random control principle are followed; CT-26 cells were cultured in RPMI 1640 medium containing 10% FBS and 1% penicillin and under cell culture conditions of 37deg.C and 5% CO 2 An incubator.
S2, 200 mu L of iron death inducer is injected into the tail of the mouse intravenously, and the injection dosage is 6mg/kg.
S3, in vivo tumor imaging, injecting 150mg of luciferase substrate into the abdominal cavity of the mouse after 7 days of treatment, performing in vivo bioluminescence imaging, obtaining a luminescent image through a spectrum imaging system Perkinelmer, detecting CT-26 cells, taking spleen and peripheral blood cells of the mouse after 21 days, performing flow cytometry analysis, and monitoring colon tumor growth condition of the mouse.
Results: referring to fig. 2, after different treatments, the fluorescence intensities of CD4 and CD8 of colon tumor tissue immune cells of the mice of the control example 2 are common, the colon tumor volume of the mice of the control example 2 is not significantly changed after 2-3 days of treatment, and the colon tumor volume of the mice of the control example 2 is not significantly different from that of the mice of the control example 1 after 20 days of treatment, which indicates that the iron death inducer of the control example 2 cannot accelerate the Fenton reaction after lacking the hot environment provided by ultrasound, so that the tumor cells cannot sensitize the iron death, and thus the tumor microenvironment cannot be effectively inhibited.
Comparative example 3
The difference from example 1 is that the FMT was not added and only the ultrasonic treatment was performed, specifically comprising the steps of:
s1, establishing a colon tumor model, subcutaneously inoculating colon tumor cells on the right side of a 4-week BALB/c female mouse, and culturing for 5 days; after the BALB/c female mice are weighed, the random groups are designed in groups, each group has 4 mice in weight in each interval, the individual biological difference among the groups of each embodiment is ensured to be minimized, and the consistency principle and the random control principle are followed; CT-26 cells were cultured in RPMI 1640 medium containing 10% FBS and 1% penicillin and under cell culture conditions of 37deg.C and 5% CO 2 An incubator.
S2, adjusting ultrasonic sound intensity, monitoring by using a thermal imager, generating heat to a tumor part by an ultrasonic probe at 43 ℃, and generating ultrasonic frequency of 1.0MHz and ultrasonic sound intensity of 0.75W/cm 2 Ultrasonic time is 10min.
S3, in vivo tumor imaging, injecting 150mg of luciferase substrate into the abdominal cavity of the mouse after 7 days of treatment, performing in vivo bioluminescence imaging, obtaining a luminescent image through a spectrum imaging system Perkinelmer, detecting CT-26 cells, taking spleen and peripheral blood cells of the mouse after 21 days, performing flow cytometry analysis, and monitoring colon tumor growth condition of the mouse.
Results: as shown in FIG. 2, after different treatments, the fluorescence intensity of the colon tumor tissue immune cells CD4 and CD8 of the mice of the control example 3 is low, and after the treatment is carried out for 2-3 days, the colon tumor volume of the mice of the control example 3 is not obviously changed, and the colon tumor cells are not effectively inhibited after the treatment is carried out for 20 days.
FIG. 3 shows the expression of CD4 and CD8 in the subcutaneous colon tumor tissue of the mice in example 1 and comparative examples 1-3, and FIG. 2 shows the fluorescence imaging of CD4 and CD8 in the subcutaneous colon tumor tissue of the mice in example 1 and comparative examples 1-3, wherein the immune cells CD4 and CD8 of the colon tumor tissue of the mice in example 1 are obviously more than those in comparative examples 1-3 after different treatments, and the fluorescence imaging effect is better than that in comparative examples 1-3;
FIG. 4 shows the change in subcutaneous colon tumor volume after treatment of mice of example 1 and comparative examples 1-3 of the present invention. It can be seen that the colon tumor volumes of the mice of examples 1 and 1-3 were not significantly changed after 2-3 days of simultaneous treatment; after 20 days of treatment, the colon tumor volume of the mice of example 1 was significantly reduced compared to the mice of comparative examples 1-3, whereas the subcutaneous colon tumor volume tended to increase regardless of PBS buffer, nano-iron oxide, or sole ultrasound treatment, indicating that colon tumor cells were not effectively inhibited.
The results of the above examples, together with the results of the control, demonstrate that if not treated effectively with an iron death inducer, growing solid tumors can lead to progressive T cell hypoplasia in the peripheral blood and spleen; whereas the method of example 1 can not only eliminate solid tumors, but also maintain cd4+ and cd8+ T cells at nearly normal levels.
Finally, GPX4 is a class of iron death core regulatory proteins that are significantly down-regulated during apoptosis of cell-derived iron; referring to FIGS. 5 and 6, in the detection of iron death-related protein GPX4 in mouse tumor cells, the relative fluorescence intensity of GPX4 in example 1 is significantly lower than that in comparative examples 1-3; it can be seen that colon tumor cells are more sensitive to the iron death inducer of example 1.
In the application process of the existing iron death inducer, the effect of inhibiting tumor by using FMT alone is poor, but the tumor cannot be treated by ultrasound alone, and in the preferred embodiment 1 of the invention, mild hyperthermia (43 ℃) triggered by ultrasound is found by using FMT and moderate ultrasonic heating, fenton reaction is accelerated in tumor cells treated by nano ferric oxide, and iron death and apoptosis are induced; whereas gentle hyperthermia reduces the risk of thermal diffusion in high intensity pulsed ultrasound therapy with great precision and widens the therapeutic indications of low intensity pulsed ultrasound.
In conclusion, the application of the iron death inducer provided by the invention can accelerate Fenton reaction in an iron-rich environment provided by nano iron oxide particles, so that colon tumor cells can sensitize iron death, and immune reaction is stimulated to overcome inherent immunity and inhibit tumor microenvironment.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (7)
1. An application of an iron death inducer, which is characterized by comprising the following steps:
adding an iron death inducer into a target sample, simultaneously carrying out ultrasonic treatment on the target sample by adopting ultrasonic waves, controlling the sound intensity and frequency of the ultrasonic waves, maintaining the temperature of the target sample at 43 ℃, and inducing the iron death of the target sample;
the target sample is an isolated tissue or a cell, the cell comprising a colon tumor cell; the iron death inducer is nano iron oxide particle FMT and is used for providing an iron environment for the target sample; the ultrasound is used to provide a thermal environment for the target sample.
2. The use of an iron death-inducing agent according to claim 1, wherein said nano iron oxide particle FMT has a concentration of 0.75-1mg/mL.
3. Use of an iron death inducer according to claim 1, wherein the nano iron oxide particle FMT has a molecular weight of 750kDa.
4. Use of an iron death inducer according to claim 1, wherein the intensity of ultrasound is between 0.75 and 1.2W/cm 2 The ultrasonic frequency was 1.0MHz.
5. The use of an iron death inducer according to claim 1, wherein the time of sonication of the target sample is between 5 and 10 minutes.
6. The use of an iron death inducer according to claim 1, wherein the culture condition of the target sample is 37 ℃,5% co 2 An incubator.
7. The use of an iron death inducer according to claim 1, wherein said colon tumour cells are CT-26 or HT-29 cells.
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